Antibodies for FFPE

Does this antibody work? Designing a new panel?
Antibodies, isotopes, etc. for Imaging Mass Cytometry applications
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ssivajothi
Posts: 33
Joined: Thu Jan 11, 2018 9:00 pm

Antibodies for FFPE

Post by ssivajothi » Tue Jan 30, 2018 7:05 pm

Hi all,

One of the main challenges I have faced in working with IMC is finding antibodies that work on FFPE tissues. There are plenty available and many have been added to the Fluidigm catalog, but you still have to spend quite a while searching the web to find validated antibodies. In order to help with this, I am attaching here the curated catalog from Cell Signaling Technologies that I have scraped from their website which includes all antibodies that should work on paraffin embedded sections. CST antibodies are very good and their validation is spot on so I have confidence that all of these antibodies should work. They do custom formulations on request so that we can get it carrier free for conjugation. If enough people express interest in this they may even be open to having conjugation-ready antibodies in the future (this is only my speculation).

If anyone else would like to share lists of clones that they have validated, I am sure it will be useful to the entire community and eliminate duplication of effort. I will do the same as I test and develop lists of working clones from multiple sources.

https://docs.google.com/spreadsheets/d/ ... sp=sharing

Best,
Santhosh Sivajothi
douglas.linn
Posts: 18
Joined: Mon Jan 22, 2018 4:20 pm

Re: Antibodies for FFPE

Post by douglas.linn » Tue Jan 30, 2018 8:48 pm

I would concur that many CST antibodies work quite well, especially for mouse FFPE tissue. I also have had good success with abcam's "IHC-P" validated antibodies, particularly the EPR clones (rabbit monoclonal).
ssivajothi
Posts: 33
Joined: Thu Jan 11, 2018 9:00 pm

Re: Antibodies for FFPE

Post by ssivajothi » Thu Feb 22, 2018 3:08 pm

Hi Douglas,

I have also tested some Abcam antibodies, mostly EPR clones. They work well, but my main issue with them is cost. Many of them run between 450-600 for carrier free formulations and researchers typically balk at these prices. Yes, CyTOF/IMC is not a 'cheap' technology but I am always on the lookout for cheaper alternatives/clones to reduce the cost for my research collaborators and increase adoption.

I have found that some FFPE validated clones are available on Biolegend as well, which is an ideal source since it is cheap, quick and almost all are carrier free. I have also found many antibodies from Thermo Fisher work well (not all though, so you'd have to test) and prices are good for 100ug quantities.

I am slowly compiling a list of clones and sources and will be able to share once I have enough entries.

Best,
Santhosh
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